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. 2018 Feb 14;8:3029. doi: 10.1038/s41598-018-21050-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Changes with time after seeding of the state of a cell culture in a traditional assay in vehicle control (0.01% DMSO) conditions. (a) Lactate (measured by LC-MS) and (b) glutamine (measured by LC-UV) in spent medium. (c) Number of live cells per well as measured using the Trypan blue exclusion technique using a Countess automated cell counter (Thermo Scientific, Loughborough, UK). Zero time corresponds to 24 hours after seeding of cells into a medium containing 10 mM of glucose, 2 mM of glutamine, in addition to dialyzed fetal calf serum, vitamins and both essential and non-essential amino acids at concentrations well below 1 mM except for arginine (0.95 mM) and glutamine (2 mM), as shown in Table 2 in Materials and Methods. The cell lines were: A549 (left) and H358 (right). For this single experiment, measurements were performed in triplicate. Shown are the mean ± SEM for the 3 technical replicates per cell line in control conditions.